CN105158125B - A kind of telomere length measuring method - Google Patents
A kind of telomere length measuring method Download PDFInfo
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- CN105158125B CN105158125B CN201510318821.4A CN201510318821A CN105158125B CN 105158125 B CN105158125 B CN 105158125B CN 201510318821 A CN201510318821 A CN 201510318821A CN 105158125 B CN105158125 B CN 105158125B
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Abstract
The invention discloses a kind of telomere length measuring method, this method need to be used to two kinds of surface-enhanced Raman scattering probe particles, a kind of telomere SERS probe particles for that can hybridize with telomeric sequence, another kind are the centromere SERS probe particles that can hybridize with centromeric sequence.Telomere SERS probes and centromere SERS probes are added on the cell sample fixed with fixative, telomere SERS probes and centromere SERS probes is hybridized to respectively by the DNA sequence dna of their surface modifications on telomere and centromere in chromosome.The relative length of telomere is characterized with the SERS signal intensity ratio of centromere SERS probes using the SERS signal intensity of telomere SERS probes.RT/CThe telomere of bigger expression cell is longer, RT/CThe telomere of smaller expression cell is shorter.Using in situ hybridization SERS method, there is the advantages of experimental cost is low, the cycle is short, specificity is good, the degree of accuracy is high.
Description
Technical field
The present invention relates to nano material preparation and biomolecule detection technology, more particularly to a kind of unicellular interior high sensitivity
Telomere length measuring method, by the spy that the high sensitivity and spectral information of SERS (SERS) are abundant
Point, realize that the telomere length in unicellular measures by hybridization in situ technique, this method is alternatively referred to as in situ hybridization surface enhanced
Raman scattering method.
Background technology
Telomere is a bit of DNA- protein complexes positioned at eukaryotic linear chromosomal end, and it is protection that it, which is acted on,
The end of chromosomal DNA, avoid degraded, terminal fusion and improper restructuring of chromosomal DNA etc..In mammalian cell
In, telomeric dna is repeated to be composed in series by TTAGGG sequences.Because problem is replicated in DNA end, each division of cell is all adjoint
The loss of a bit of sequence of telomeric dna.Each division of cell all can be along with the shortening of end of chromosome telomere, cell
Therefore gradually aging, until lose multiplication capacity and it is dead.The change of telomere length and a variety of diseases are closely related, as cancer,
Early ageing syndrome etc..Therefore measurement telomere length can provide the important information related to disease.
Current existing telomere length measuring method includes：Southern blottings, hybridize Protection Code (HPA), be glimmering in real time
Fluorescent Quantitative PCR method (qPCR) and FISH (FISH) method.DNA with Southern Northern blot analysis is not broken
It is higher with purity, but the data that this method obtains further comprises part subtelomeric section length, therefore Southern blottings are not
The physical length of telomere can be provided.In HPA methods, directly genomic DNA and the specific acridinium ester probe of telomere are hybridized, passed through
Chemiluminescence determination telomere length.Radio isotope need not be used in this method, is greatly reduced to experimental implementation people
The injury of member, while there is no rigors to genomic integrity and purity.But HPA methods do not provide detailed cell or dyeing
The horizontal telomere information of body, can not directly determine telomere length.PCR methods use the mode of PCR amplifications, effectively reduce cell
The dosage of genomic DNA.FISH methods utilize telomere-specific peptide nucleic acid fluorescent probe, with reference to fluorescence microscopy, can realize
The Telomere length assay of individual cell level.Although these methods telomere research in be made that important contribution, they or it is more
Or there are some shortcomings less, for example sensitivity is low, program is complicated, cost is high etc., therefore, still need to design at present high sensitivity,
The telomere length e measurement technology that accuracy is good, applicability is extensive, workable.
In biological study, in addition to traditional biological detection method, optical detective technology with its it is simple, reliable the characteristics of obtain
Obtained relatively broad application.Existing optical test method obtains relatively broad application as main flow using fluorescent technique.Make
For another spectral technique, Raman spectrum carries structure " fingerprint " information of tested molecule, thus has prior biology
Learn application value.But because its spectral signal intensity is too weak, limit its application.SERS (SERS) phenomenon
Discovery thoroughly solve this problem so that it turns into a kind of strong technical Analysis means.SERS on the one hand after
The plurality of advantages of Raman spectrum has been held, small, spectral information such as is damaged to biological tissue and enriched；On the other hand, it compensate for passing
System Raman scattering signal intensity is weak, is unfavorable for the shortcomings that detection.Its huge humidification causes the spectral detection based on SERS
Sensitivity with superelevation, or even the analysis and research of single molecules level can be realized." fingerprint " characteristic of SERS spectra allows one to
Tracking, detection target molecule in the biotic environment of complexity.SERS effects are produced in the coarse metal surface of nanoscale, nanometer
The rapid development of technology provides abundant technological approaches to construct the SERS of multifunction nano-probes/substrate.These are based on
Nano-probe/substrate of SERS spectra technology is many in bio-imaging, nucleic acid or Protein Detection, tumour identification, drug delivery etc.
Biomedical sector has shown excellent application prospect.
The content of the invention
Goal of the invention：To solve the problems, such as that not high current telomere length measuring method sensitivity, complex operation, cost are high,
The present invention provides a kind of in situ hybridization SERS methods for the measurement of cell telomere length, by SERS high sensitivity and spectrum
The characteristics of abundant information, realize that the telomere length in unicellular measures by hybridization in situ technique.
Technical scheme：To achieve the above object, the technical solution adopted by the present invention is：
A kind of telomere length measuring method, marked respectively using telomere SERS probe particles and centromere SERS probe particles
The telomere of cell to be measured and centromere, and it is strong using the SERS signal of telomere SERS probe particles and centromere SERS probe particles
The ratio between degree RT/CThe telomere length of cell to be measured is characterized, i.e., keeps constant telomeric sequence to be used as internal reference, elimination using length
The influence of the concentration of SERS probes, chromosome or cell；This method is specially：By telomere SERS probe particles and centromere SERS
Probe particle is added on the cell to be measured fixed with fixative, makes telomere SERS probe particles and centromere SERS probe grains
Son is hybridized on the telomere and centromere of cell to be measured by the DNA sequence dna of their surface modifications respectively, is visited using telomere SERS
The SERS signal intensity of pin particle and the SERS signal intensity ratio R of centromere SERS probe particlesT/CCharacterize the end of cell to be measured
Grain length, RT/CThe telomere length of smaller expression cell to be measured is shorter；The telomere SERS probe particles be it is a kind of can be with cell
Telomeric sequence hybridization SERS probe particles, the centromere SERS probe particles be it is a kind of can be miscellaneous with the centromeric sequence of cell
The SERS probe particles of friendship.
The above method specifically comprises the following steps：
Step 1：Prepare color of spherical gold A；
Step 2：In the complementary DNA sequence dna 5'-SH (CH of the telomere of particle A surface modifications and cell2)6TTTTTTTTTTCCCTAACCCTAACCCTAA-3', obtain the golden nanometer particle B that can hybridize with the telomeric sequence of cell1；In grain
The complementary DNA sequence dna 5'-SH (CH in the centromere of sub- A surface modifications and cell2)6TTTTTTTTTTTAGCTTCTGTCTAGTTT-
3', obtain the golden nanometer particle B that can hybridize with the centromeric sequence of cell2；
Step 3：In particle B1Raman reporter molecules are modified on the room on surface, obtain producing SERS signal and can be with
The golden nanometer particle C of the telomeric sequence hybridization of cell1；In particle B2Raman reporter molecules are modified on the room on surface, obtain producing
Raw SERS signal and the golden nanometer particle C that can hybridize with the centromeric sequence of cell2；It is required that in particle B1With particle B2Repair on surface
The Raman reporter molecules of decorations are different so that particle C1With particle C2Caused SERS signal is different；
Step 4：In particle C1One layer of silver-colored hull shape of Surface coating is into telomere SERS probe particles D1, silver-colored shell is by particle D1On
Section of DNA sequence (i.e. SH (the CH of Raman reporter molecules and rich T bases2)6TTTTTTTTTTT in) being embedded in, and it is used for what is hybridized
Section of DNA sequence (i.e. CCCTAACCCTAACCCTAA) is still outside silver-colored shell；In particle C2One layer of silver-colored hull shape of Surface coating
Into centromere SERS probe particles D2, silver-colored shell is by particle D2On Raman reporter molecules and rich T bases section of DNA sequence (i.e. SH
(CH2)6TTTTTTTTTTT in) being embedded in, and the section of DNA sequence (i.e. AGCTTCTGTCTAGTTT) for being used to hybridize still exposes
Outside silver-colored shell；The thickness of silver-colored shell is 2~3nm (thickness may not exceed 3.4nm), and SERS signal is further enhanced by silver-colored shell, but
The hybridization function of DNA sequence dna is not influenceed again simultaneously；
Step 5：Cell to be measured is fixed with fixative；
Step 6：By particle D1With particle D2It is added to simultaneously in the cell to be measured fixed with fixative, makes particle D1With
Particle D2Hybridized to respectively by the DNA sequence dna of their surface modifications on the telomere and centromere of cell to be measured；
Step 7：Wash away particle D unnecessary on cell to be measured1With particle D2；
Step 8：Analyze the SERS spectra signal on cell to be measured using confocal Raman microscope, carry out SERS spectra and
The collection of image；
Step 9：Believed using the SERS signal intensity of telomere SERS probe particles and the SERS of centromere SERS probe particles
Number intensity ratio RT/CCharacterize the telomere length of cell to be measured, RT/CThe telomere length of smaller expression cell to be measured is shorter.
In the step 2, DNA sequence dna 5'-SH (CH2)6TTTTTTTTTTCCCTAACCCTAACCCTAA-3' and DNA sequences
Arrange 5'-SH (CH2)6Sulfydryl has been modified at TTTTTTTTTTTAGCTTCTGTCTAGTTT-3' 5' ends, to cause two kinds of DNA sequences
Row can by golden sulfide linkage securely, be covalently linked to particle A surface.
In the step 3, two kinds of Raman reporter molecules contain sulfydryl, to enable Raman reporter molecules to pass through golden sulphur
Key securely, be covalently linked to particle A surface；It is required that the raman scattering cross section of the two kinds of Raman reporter molecules used compared with
Greatly, it is easy to produce strong SERS signal.
Beneficial effect：Telomere length measuring method provided by the invention, relative to prior art, there is following advantage：1、
Probe particle in the present invention can provide SERS signal, have the sensitivity of superelevation, the analysis and research available for individual cell level；
2nd, internal reference is used as using the constant centromere of length in the present invention, can effectively avoid probe, cell, dye bulk concentration etc. because
The influence of element, improve the accuracy of telomere length measurement；3rd, the present invention realizes that telomere length is surveyed using SERS Probe In Situ Hybridizations
Amount, there is the advantages of experimental cost is low, the cycle is short, specificity is good, accurate positioning.
Brief description of the drawings
Fig. 1 is the schematic diagram that in situ hybridization SERS methods proposed by the present invention measure telomere length；
Fig. 2 is the structural representation of the SERS probes used in the present invention；
Fig. 3 is the SERS spectra of telomere SERS probes in embodiment 1；
Fig. 4 is the SERS spectra of centromere SERS probes in embodiment 1.
Embodiment
The present invention is further described below in conjunction with the accompanying drawings.
It is a kind of telomere length measuring method as shown in Figure 1, by telomere SERS probe particles and centromere SERS probe grains
Son is added on the cell to be measured fixed with fixative, distinguishes telomere SERS probe particles and centromere SERS probe particles
Hybridized to by the DNA sequence dna of their surface modifications on the telomere and centromere of cell to be measured, utilize telomere SERS probe particles
SERS signal intensity and centromere SERS probe particles SERS signal intensity ratio RT/CCharacterize the telomere length of cell to be measured
Degree, RT/CThe telomere length of smaller expression cell to be measured is shorter；As shown in Fig. 2 the telomere SERS probe particles be one kind can be with
Cell telomeric sequence hybridization SERS probe particles, the centromere SERS probe particles be it is a kind of can be with the centromere of cell
The SERS probe particles of sequence hybridization.This method comprises the following steps：
Step 1：Prepare color of spherical gold A；
Step 2：In the complementary DNA sequence dna 5'-SH (CH of the telomere of particle A surface modifications and cell2)6TTTTTTTTTTCCCTAACCCTAACCCTAA-3', obtain the golden nanometer particle B that can hybridize with the telomeric sequence of cell1；In grain
The complementary DNA sequence dna 5'-SH (CH in the centromere of sub- A surface modifications and cell2)6TTTTTTTTTTTAGCTTCTGTCTAGTTT-
3', obtain the golden nanometer particle B that can hybridize with the centromeric sequence of cell2；Sulfydryl has been modified at the 5' ends of two kinds of DNA sequence dnas,
With enable two kinds of DNA sequence dnas by golden sulfide linkage securely, be covalently linked to particle A surface；
Step 3：In particle B1Raman reporter molecules are modified on the room on surface, obtain producing SERS signal and can be with
The golden nanometer particle C of the telomeric sequence hybridization of cell1；In particle B2Raman reporter molecules are modified on the room on surface, obtain producing
Raw SERS signal and the golden nanometer particle C that can hybridize with the centromeric sequence of cell2；It is required that in particle B1With particle B2Repair on surface
The Raman reporter molecules of decorations are different so that particle C1With particle C2Caused SERS signal is different；Two kinds of Raman reporter molecules contain
Sulfydryl, with enable Raman reporter molecules by golden sulfide linkage securely, be covalently linked to particle A surface.
Step 4：In particle C1One layer of silver-colored hull shape of Surface coating is into telomere SERS probe particles D1, silver-colored shell is by particle D1On
In the section of DNA sequence of Raman reporter molecules and rich T bases is embedded in, and the section of DNA sequence for being used to hybridize still is exposed to
Outside silver-colored shell；In particle C2One layer of silver-colored hull shape of Surface coating is into centromere SERS probe particles D2, silver-colored shell is by particle D2On Raman report
In the section of DNA sequence of announcement molecule and rich T bases is embedded in, and the section of DNA sequence for being used to hybridize still is exposed to outside silver-colored shell；
The thickness of silver-colored shell is 2~3nm, and SERS signal is further enhanced by silver-colored shell, but simultaneously and does not influence the hybridization work(of DNA sequence dna
Energy；
Step 5：Cell to be measured is fixed with fixative；
Step 6：By particle D1With particle D2It is added to simultaneously in the cell to be measured fixed with fixative, makes particle D1With
Particle D2Hybridized to respectively by the DNA sequence dna of their surface modifications on the telomere and centromere of cell to be measured；
Step 7：Wash away particle D unnecessary on cell to be measured1With particle D2；
Step 8：Analyze the SERS spectra signal on cell to be measured using confocal Raman microscope, carry out SERS spectra and
The collection of image；
Step 9：Believed using the SERS signal intensity of telomere SERS probe particles and the SERS of centromere SERS probe particles
Number intensity ratio RT/CCharacterize the telomere length of cell to be measured, RT/CThe telomere length of smaller expression cell to be measured is shorter.
The present invention is further illustrated with reference to embodiment.
Embodiment 1：
With the Raman reporter molecules that 3,5- thiophenol dichlorobenzenes (3,5-DCT) molecule is centromere SERS probe particles, with 5,
The thiobis of 5'- bis- (2- nitrobenzoic acids) molecule is the Raman reporter molecules of telomere SERS probe particles, thin with human cervical carcinoma
Born of the same parents (HeLa) are cell to be measured, and telomere length measurement is carried out using the method for the present invention.
1) color of spherical gold is prepared
200 μ L 10%HAuCl are added in 200mL deionized waters4Solution, it is stirred vigorously and is heated to seething with excitement.Then add
Enter the sodium citrate aqueous solutions of 8mL 1%, continue heating stirring 15min.Stop heating, stir to solution and be cooled to room temperature, produce
To the solution of gold nanoparticles of claret.
2) SERS probe particles are prepared
Telomere SERS probe particles：140 μ L10 μM are added in 1mL color of spherical gold solution and are dissolved in PBS bufferings
DNA sequence dna (5'-SH (CH in liquid2)6TTTTTTTTTTCCCTAACCCTAACCCTAA-3'), the mixed solution is stored at room temperature
After 12h, 1 μ L 10mM DTNB solution is added, the mixed solution continues to stand 6h, and then adding 100 μ L every 1h contains 0.3M
NaCl and 0.1% lauryl sodium sulfate (SDS) PBS, add 5 times altogether.The mixed liquor continues to stand 12h, then uses
7500rpm, 30min eccentric cleaning 1 time, precipitation are dispersed in 1mL deionized waters.
The silver nitrate that 10 μ L 10mM are added in the nano-particle that obtained 1mL is connected with DNA sequence dna and DTNB molecules is molten
Liquid, wraps up silver-colored shell, and stirring adds 22 μ L 10mM ascorbic acid solution reaction 6h after 30 minutes, then with 7000rpm,
30min eccentric cleanings 1 time, precipitation are dispersed in the PBS that 1mL contains 0.4 μ g/mL salmon sperm dnas, and 4 DEG C of preservations are stand-by,
Obtain telomere SERS probes.
The preparation of centromere SERS probe particles is similar with telomere SERS probe particles, and DNA sequence dna only is replaced with into (5'-
SH(CH2)6TTTTTTTTTTTAGCTTCTGTCTAGTTT-3'), DTNB is replaced with 3,5-DCT, remaining step dosage is identical.
Obtain centromere SERS probe particles.
3) in situ hybridization cell sample prepares
HeLa cells are inoculated in 35mm Glass bottom culture dish, cultivated 24 hours.Fixed carefully with 4% paraformaldehyde solution room temperature
Born of the same parents 20 minutes.Gently rinsed twice with PBS.Add the μ g/mL RNase of 1mL 100 and be incubated 20 minutes in 37 DEG C, used
PBS is gently rinsed twice.Add the pepsins of 2mL 0.005% (solvent is 10mM HCl, and 45 DEG C are preheated to before use) and in
37 DEG C are incubated 5 minutes, are gently rinsed twice with PBS.It is molten that PBSs of the 2mL containing 0.4 μ g/mL salmon sperm dnas is added into two samples
4 DEG C of closing 4h of liquid, are then gently rinsed 3 times with PBS.
5) SERS Probe In Situ Hybridizations
200 μ L telomere SERS probe particles and centromere SERS probe particles is respectively taken to be added to 1.6mL and contain 0.4 μ g/mL salmons
It is well mixed in the PBS solution of milt DNA, is preheated 5 minutes in 85 DEG C.2mL is separately added into two Glass bottom culture dish samples
Preheated SERS probes, make its uniform fold glass bottom.After 85 DEG C are heated 10 minutes, room temperature hybridizes 6 hours.With cleaning solution (2
× SSC, 0.1% polysorbas20) room temperature cleaning twice, every time cleaning 15 minutes, to remove non-hybridized SERS probe particles.Go from
Sub- water cleaning is once afterwards dried up Glass bottom culture dish with argon gas.
6) telomere length measures
Intracellular centromere SERS probes and telomere are gathered using the confocal Raman microscope with mapping functions
The ratio between the SERS spectra and image of SERS probes, and analyze the SERS signal of telomere SERS probes and centromere SERS probes, obtain
Take the information of telomere relative length.
The SERS spectras of the telomere SERS probes prepared in the embodiment is as shown in figure 3, centromere SERS probes
SERS spectra is as shown in Figure 4.
Embodiment 2：
Telomere SERS probe particles and centromere SERS probe particles are prepared according to the method in embodiment 1.Use HeLa
Cell and MRC-5 cells, cell sample is prepared according to the method in embodiment 1.Carried out according to the method in embodiment 1 in situ miscellaneous
Friendship and the collection of SERS spectra and image.Analysis result discovery, the R of MRC-5 cellsT/CMore than the R of HeLa cellsT/C, explanation
The telomere of MRC-5 cells is longer than the telomere of HeLa cell.
Embodiment 3：
Telomere SERS probe particles and centromere SERS probe particles are prepared according to the method in embodiment 1.Use HeLa
Cell is tested.One group of HeLa cell co-cultures 2 weeks with 0.8mM telomerase inhibitors retrovir (AZT), and one group of HeLa is thin
Born of the same parents do not co-culture with telomerase inhibitor.Prepare cell sample according to the method in embodiment 1 and carry out in situ hybridization and
The collection of SERS spectra and image.Analysis result find, not with AZT co-culture HeLa cells RT/CCo-cultured more than with AZT
HeLa cells RT/C, illustrate that the former telomere is longer than the latter, i.e. AZT restrained effectively the function of Telomerase so as to yes
The telomere of HeLa cells shortens.
Described above is only the preferred embodiment of the present invention, it should be pointed out that：For the ordinary skill people of the art
For member, under the premise without departing from the principles of the invention, some improvements and modifications can also be made, these improvements and modifications also should
It is considered as protection scope of the present invention.
SEQUENCE LISTING
<110>Southeast China University
<120>A kind of telomere length measuring method
<130> 2015
<160> 2
<170> PatentIn version 3.3
<210> 1
<211> 28
<212> DNA
<213>Artificial sequence
<223>The complementary DNA sequence dna of telomere
<400> 1
tttttttttt ccctaaccct aaccctaa 28
<210> 2
<211> 27
<212> DNA
<213>Artificial sequence
<223>The complementary DNA sequence dna of telomere
<400> 2
tttttttttt tagcttctgt ctagttt 27
Claims (3)
- A kind of 1. telomere length measuring method, it is characterised in that：Realize that telomere length measures using in situ hybridization SERS methods, by end Grain SERS probe particles and centromere SERS probe particles are added on the cell to be measured fixed with fixative, make telomere SERS Probe particle and centromere SERS probe particles pass through the DNA sequence dna in situ hybridization of their surface modifications to cell to be measured respectively On telomere and centromere, believed using the SERS signal intensity of telomere SERS probe particles and the SERS of centromere SERS probe particles Number intensity ratio RT/CCharacterize the telomere length of cell to be measured, RT/CThe telomere length of smaller expression cell to be measured is shorter；The end Grain SERS probe particles are a kind of SERS probe particles that can hybridize with the telomeric sequence of cell, the centromere SERS probe grains Son is a kind of SERS probe particles that can hybridize with the centromeric sequence of cell；Method comprises the following steps：Step 1：Prepare color of spherical gold A；Step 2：In the complementary DNA sequence dna 5'-SH (CH of the telomere of particle A surface modifications and cell2)6TTTTTTTTTTCCCTAACCCTAACCCTAA-3', obtain the golden nanometer particle B that can hybridize with the telomeric sequence of cell1；In grain The complementary DNA sequence dna 5'-SH (CH in the centromere of sub- A surface modifications and cell2)6TTTTTTTTTTTAGCTTCTGTCTAGTTT- 3', obtain the golden nanometer particle B that can hybridize with the centromeric sequence of cell2；Step 3：In particle B1Raman reporter molecules are modified on the room on surface, obtain that SERS signal can be produced and can be with cell The golden nanometer particle C of telomeric sequence hybridization1；In particle B2Raman reporter molecules are modified on the room on surface, obtain that SERS can be produced Signal and the golden nanometer particle C that can hybridize with the centromeric sequence of cell2；It is required that in particle B1With particle B2The drawing of surface modification Graceful report molecule is different so that particle C1With particle C2Caused SERS signal is different；Step 4：In particle C1One layer of silver-colored hull shape of Surface coating is into telomere SERS probe particles D1, silver-colored shell is by particle D1On Raman In the section of DNA sequence of report molecule and rich T bases is embedded in, and the section of DNA sequence for being used to hybridize still is exposed to silver-colored shell Outside；In particle C2One layer of silver-colored hull shape of Surface coating is into centromere SERS probe particles D2, silver-colored shell is by particle D2On Raman reporter point In the section of DNA sequence of sub and rich T bases is embedded in, and the section of DNA sequence for being used to hybridize still is exposed to outside silver-colored shell；Silver-colored shell Thickness be 2~3nm, SERS signal is further enhanced by silver-colored shell, but simultaneously and do not influence the hybridization function of DNA sequence dna；Step 5：Cell to be measured is fixed with fixative；Step 6：By particle D1With particle D2It is added to simultaneously in the cell to be measured fixed with fixative, makes particle D1And particle D2Hybridized to respectively by the DNA sequence dna of their surface modifications on the telomere and centromere of cell to be measured；Step 7：Wash away particle D unnecessary on cell to be measured1With particle D2；Step 8：The SERS spectra signal on cell to be measured is analyzed using confocal Raman microscope, carries out SERS spectra and image Collection；Step 9：It is strong using the SERS signal intensity and the SERS signal of centromere SERS probe particles of telomere SERS probe particles The ratio between degree RT/CCharacterize the telomere length of cell to be measured, RT/CThe telomere length of smaller expression cell to be measured is shorter.
- 2. telomere length measuring method according to claim 1, it is characterised in that：In the step 2, DNA sequence dna 5'- SH(CH2)6TTTTTTTTTTCCCTAACCCTAACCCTAA-3' and DNA sequence dna 5'-SH (CH2)6Sulfydryl has been modified at TTTTTTTTTTTAGCTTCTGTCTAGTTT-3' 5' ends, to enable two kinds of DNA sequence dnas to pass through gold Sulfide linkage securely, be covalently linked to particle A surface.
- 3. telomere length measuring method according to claim 1, it is characterised in that：In the step 3, two kinds of Raman reports Accuse molecule and contain sulfydryl, with enable Raman reporter molecules by golden sulfide linkage securely, be covalently linked to particle A table Face.
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| CN105950760A (en) * | 2016-06-21 | 2016-09-21 | 南开大学 | Method for simultaneously detecting unicell mRNA (messenger ribonucleic acid) expression level and telomere length |
| CN108913754A (en) * | 2018-07-26 | 2018-11-30 | 苏州呼呼健康科技有限公司 | Detection reagent, kit and the application of sperm telomere length and detection method |
| CN112592963A (en) * | 2021-01-04 | 2021-04-02 | 东南大学 | Telomere and centromere super-resolution imaging method and probe thereof |
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|---|
| Assessing Telomere Length Using Surface Enhanced Raman Scattering;Shenfei Zong et al.;《Scientific Reports》;20141110;第4卷;第1-4、7页 * |
| Quantifying Telomere Lengths of Human Individual Chromosome Arms by Centromere-Calibrated Fluorescence in Situ Hybridization and Digital Imaging;Sven Perner et al.;《American Journal of Pathology》;20031130;第163卷(第5期);第1751-1756页 * |
| Trace analysis of mercury(II) ions using aptamer-modified Au/Ag core–shell nanoparticles and SERS spectroscopy in a microdroplet channel;Eunsu Chung et al.;《Lab on a Chip》;20131231;第13卷(第2期);第260-266页 * |
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